HUANG Qian-Hua, ZHANG Chu-Yu, WANG Ning, FU Lie-Zhen and WANG Jia-Fu. Sequencing and Com parison of NS2-3 Gene Fragm ent of Classieal Swine Fever Virus Shimen Strain[J]. Virologica Sinica, 1999, 14(2): 163-168.
Citation: HUANG Qian-Hua, ZHANG Chu-Yu, WANG Ning, FU Lie-Zhen, WANG Jia-Fu. Sequencing and Com parison of NS2-3 Gene Fragm ent of Classieal Swine Fever Virus Shimen Strain .VIROLOGICA SINICA, 1999, 14(2) : 163-168.

猪瘟病毒石门株NS23基因片段的序列测定及比较

  • 根据猪瘟病毒C株的序列,以计算机辅助设计,化学合成1对引物(PF5648/PR6604),应用RTPCR技术从感染猪血中成功地扩增了我国猪瘟病毒强毒石门株NS23基因片段,大小为957bp,位于NS3基因的中部NTPase和Helicase活性区。克隆后测序,结果表明该段基因产物具有解旋酶超家族全部七个特征性保守序列,包括共同的NTP结合基序A位点(GXGKT/S)和B位点(3hy,2x)D。序列同源性比较表明,石门株与日本的ALD和GPE-株同源性最高,与其它3株猪瘟病毒(C株、Brescia株和Alfort株)的同源性也很高,并与2株牛病毒性腹泻病毒(BVDV)(NADL株和SD1株)也有较高的同源性,尤其是由核苷酸序列推导的氨基酸序列,同源性均大于90%,是瘟病毒属基因组中最保守的区段,这与该基因产物在病毒复制及聚蛋白前体加工过程中所具有的重要功能是一致的

Sequencing and Com parison of NS2-3 Gene Fragm ent of Classieal Swine Fever Virus Shimen Strain

  • A couple of CSFV specific primer(PF 5648 and PR 6604)was designed with the aid of computer primer designation software and synthesized based upon the relative conserved regions of published sequenL-~s of C strain.NS2-3 gene(p125 gene)fragment of CSFV Shimen strain was amplified successfu~y by RT-PCR from the anticoagulant blood of infected pig. The product length is 957 bp,located in the central of NS3,a putative NTPase and helicase domain.The ob tained PCR product was cloned and then sequenced.The sequence showed that this fragment con tained all of seven typical consexved segments of helicase superfamily, including two common N11Pbinding motifs,namely,Asite(GXGKT/S)andBsite(3hy,2x)D.Sequence ho mology analysis revealed that Shimen strain had the highest homology wi th Japanese strains(ALD and GPE ),and slightly lower homology with other three CSFV strains (C.Brescia and AI fort).Shimen strain had also significant homology with two BVDV strains (NADL and SD-1) The deduced amino acid sequence homology of Shimen strain with five CSFV and two BVDV strains was all upper than 90% .It is further confirmed that this fragment is the most co nserved in pestivirns amino acid sequence It is cons istent with its essential function in replication and trans lation of virus genome and in processing of polypmtein precursor

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    Sequencing and Com parison of NS2-3 Gene Fragm ent of Classieal Swine Fever Virus Shimen Strain

    • 1. Institute of Virology,Wuhan University,Wuhan 430072

    Abstract: A couple of CSFV specific primer(PF 5648 and PR 6604)was designed with the aid of computer primer designation software and synthesized based upon the relative conserved regions of published sequenL-~s of C strain.NS2-3 gene(p125 gene)fragment of CSFV Shimen strain was amplified successfu~y by RT-PCR from the anticoagulant blood of infected pig. The product length is 957 bp,located in the central of NS3,a putative NTPase and helicase domain.The ob tained PCR product was cloned and then sequenced.The sequence showed that this fragment con tained all of seven typical consexved segments of helicase superfamily, including two common N11Pbinding motifs,namely,Asite(GXGKT/S)andBsite(3hy,2x)D.Sequence ho mology analysis revealed that Shimen strain had the highest homology wi th Japanese strains(ALD and GPE ),and slightly lower homology with other three CSFV strains (C.Brescia and AI fort).Shimen strain had also significant homology with two BVDV strains (NADL and SD-1) The deduced amino acid sequence homology of Shimen strain with five CSFV and two BVDV strains was all upper than 90% .It is further confirmed that this fragment is the most co nserved in pestivirns amino acid sequence It is cons istent with its essential function in replication and trans lation of virus genome and in processing of polypmtein precursor

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